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Cell biology assays

Secreted alkaline phosphatase assays

The Great EscAPe SEAP kits use a secreted alkaline phosphatase assay to analyze cis-acting DNA sequences and trans-acting factors. The kinetics of gene expression can easily be studied with this secreted alkaline phosphatase assay by repeatedly sampling the same culture. The kit has an extremely low background signal, with a wide dynamic range and high sensitivity. Furthermore, the robust signal is extremely stable over time. Cells can be studied further after the secreted alkaline phosphatase assay using northern blots, RNase protection assays, western blots, and other methods.

Fluorescence-based detection

The Great EscAPe SEAP Fluorescence Detection Kit uses the fluorescent substrate 4-methylumbelliferyl phosphate (MUP) and has sensitivity comparable to firefly luciferase assays. Like the chemiluminescence assay, the secreted alkaline phosphatase fluorescence assay is linear over a 10⁴-fold range of enzyme concentrations.

Dual secreted reporter assay—SEAP & luciferase

The Ready-To-Glow Dual Secreted Reporter Assay has two live cell reporters that do not require cell lysis. In addition to secreted alkaline phosphatase, it includes Metridia luciferase, which has a 2–4 fold higher signal compared to Renilla and firefly luciferases.

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Cat. #	Product	Size
631738	Great EscAPe™ SEAP Chemiluminescence Kit 2.0	1,000 Rxns	USD $1049.00
The Great EscAPe SEAP Chemiluminescence Kit 2.0 is a complete system for the detection of secreted alkaline phosphatase (SEAP) activity in the growth media from mammalian cells expressing this enzyme. The kit contains reagents necessary for the chemiluminescent detection of SEAP activity using a 96-well plate luminometer. The assay can also be performed in single tubes and measured in a tube luminometer. It allows you to monitor promoter activity over time, because samples of the cell media can be taken repeatedly and tested without lysing cells. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back SEAP activity is highly specific, with a >20-fold increase in activity SEAP activity is highly specific, with a >20-fold increase in activity. HEK 293 cells were transiently transfected with a promoter construct containing CRE driving the expression of SEAP, or mock-transfected. 12 hr posttransfection, the cell culture media was replaced by media containing either 10 μM forskolin or plain cell culture media and incubated for 7 hr. Forskolin causes an increase in the level of cytosolic cAMP, which in turn activates CRE, driving the expression of SEAP, which is detectable in the media culture supernatant. The samples were assayed using the Great EscAPe SEAP Chemiluminescence Kit 2.0 and its protocol and analyzed on a BD Monolight 3096 Luminometer. Back Comparison of the sensitivities of SEAP and firefly luciferase Comparison of the sensitivities of SEAP and firefly luciferase. Parallel cultures of BHK cells were transiently transfected with the indicated amounts of either pSEAP2-Control Vector or a similar vector expressing luciferase. After 24 hr, SEAP activity was assayed in the appropriate culture media using the Great EscAPe chemiluminescent assay. Similarly, after 24 hr, cell lysates were prepared from the luciferase cultures, and luciferase activity was assayed using a commercial kit. RLU = relative light units. Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription.** Back 631738: Great EscAPe SEAP Chemiluminescence Kit 2.0
631737	Great EscAPe™ SEAP Chemiluminescence Kit 2.0	300 Rxns	USD $588.00
The Great EscAPe SEAP Chemiluminescence Kit 2.0 is a complete system for the detection of secreted alkaline phosphatase (SEAP) activity in the growth media from mammalian cells expressing this enzyme. The kit contains reagents necessary for the chemiluminescent detection of SEAP activity using a 96-well plate luminometer. The assay can also be performed in single tubes and measured in a tube luminometer. It allows you to monitor promoter activity over time, because samples of the cell media can be taken repeatedly and tested without lysing cells. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back SEAP activity is highly specific, with a >20-fold increase in activity SEAP activity is highly specific, with a >20-fold increase in activity. HEK 293 cells were transiently transfected with a promoter construct containing CRE driving the expression of SEAP, or mock-transfected. 12 hr posttransfection, the cell culture media was replaced by media containing either 10 μM forskolin or plain cell culture media and incubated for 7 hr. Forskolin causes an increase in the level of cytosolic cAMP, which in turn activates CRE, driving the expression of SEAP, which is detectable in the media culture supernatant. The samples were assayed using the Great EscAPe SEAP Chemiluminescence Kit 2.0 and its protocol and analyzed on a BD Monolight 3096 Luminometer. Back Comparison of the sensitivities of SEAP and firefly luciferase Comparison of the sensitivities of SEAP and firefly luciferase. Parallel cultures of BHK cells were transiently transfected with the indicated amounts of either pSEAP2-Control Vector or a similar vector expressing luciferase. After 24 hr, SEAP activity was assayed in the appropriate culture media using the Great EscAPe chemiluminescent assay. Similarly, after 24 hr, cell lysates were prepared from the luciferase cultures, and luciferase activity was assayed using a commercial kit. RLU = relative light units. Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription.** Back 631737: Great EscAPe SEAP Chemiluminescence Kit 2.0
631736	Great EscAPe™ SEAP Chemiluminescence Kit 2.0	50 Rxns	USD $261.00
The Great EscAPe SEAP Chemiluminescence Kit 2.0 is a complete system for the detection of secreted alkaline phosphatase (SEAP) activity in the growth media from mammalian cells expressing this enzyme. The kit contains reagents necessary for the chemiluminescent detection of SEAP activity using a 96-well plate luminometer. The assay can also be performed in single tubes and measured in a tube luminometer. It allows you to monitor promoter activity over time, because samples of the cell media can be taken repeatedly and tested without lysing cells. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Chemiluminescent detection of protein-protein interactions Chemiluminescent detection of protein-protein interactions. The bait vector pM-53 expresses p53 fused to the GAL4 DNA-binding domain. When HEK 293 cells were cotransfected with this bait together with the SV40 large T antigen prey vector, pVP16-T, strong expression of SEAP was detected because large T antigen interacts with p53. The CP protein, which does not interact strongly with p53, was used as a negative control. Back SEAP activity is highly specific, with a >20-fold increase in activity SEAP activity is highly specific, with a >20-fold increase in activity. HEK 293 cells were transiently transfected with a promoter construct containing CRE driving the expression of SEAP, or mock-transfected. 12 hr posttransfection, the cell culture media was replaced by media containing either 10 μM forskolin or plain cell culture media and incubated for 7 hr. Forskolin causes an increase in the level of cytosolic cAMP, which in turn activates CRE, driving the expression of SEAP, which is detectable in the media culture supernatant. The samples were assayed using the Great EscAPe SEAP Chemiluminescence Kit 2.0 and its protocol and analyzed on a BD Monolight 3096 Luminometer. Back Comparison of the sensitivities of SEAP and firefly luciferase Comparison of the sensitivities of SEAP and firefly luciferase. Parallel cultures of BHK cells were transiently transfected with the indicated amounts of either pSEAP2-Control Vector or a similar vector expressing luciferase. After 24 hr, SEAP activity was assayed in the appropriate culture media using the Great EscAPe chemiluminescent assay. Similarly, after 24 hr, cell lysates were prepared from the luciferase cultures, and luciferase activity was assayed using a commercial kit. RLU = relative light units. Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription.** Back 631736: Great EscAPe SEAP Chemiluminescence Kit 2.0
631734	Ready-To-Glow™ Dual Secreted Reporter Assay	500 Rxns	USD $827.00
The Ready-To-Glow Dual Secreted Reporter Assay contains the buffers and substrates for use with the vectors contained in the Ready-To-Glow Dual Secreted Reporter Vector Kit (Cat. No. 631735). This assay allows you to monitor the activity of two promoters simultaneously. The promoters are monitored by testing the media from cells that have been transfected with the two reporter vectors, which express luciferase and SEAP respectively. The assay allows you to monitor promoter activity over time, because samples of the cell media can be taken repeatedly and tested without lysing cells. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components You May Also Like Image Data Resources Back Similar secretion kinetics of Metridia secreted luciferase and SEAP enable accurate comparisons of the relative timing of promoter activity *Similar secretion kinetics of Metridia* secreted luciferase and SEAP enable accurate comparisons of the relative timing of promoter activity. HeLa cells were plated into 6-well plates and transiently transfected with either the pMetLuc-Control vector or the pSEAP-Control vector. Media samples from the transfected cells were collected from 3 wells at each time point by removing enough media to run either the luciferase or the SEAP assay. Each sample was tested in triplicate, using a white-bottom 96-well microtiter plate on a Turner BioSystems Veritas Luminometer. Back Monitoring activation of two promoters simultaneously Monitoring activation of two promoters simultaneously.** HEK 293 cells cotransfected with pNF kappa B-TA-MetLuc and pCRE-SEAP constructs were treated with fresh media alone or with media containing either 1,000 ng/ml TNF-alpha or 10 µM forskolin. Samples of culture supernatant were collected 7 hr later and assayed using a BD Monolight 3096 Luminometer. Back 631734: Ready-To-Glow Dual Secreted Reporter Assay
631717	pSEAP2-Control Vector	20 ug	USD $568.00
pSEAP2-Control is designed to provide a reference for comparing the activities of promoters and enhancers using the secreted form of human alkaline phosphatase (SEAP) as a reporter gene. This vector can also be used to monitor transfection efficiencies. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back pSEAP mammalian reporter vectors pSEAP mammalian reporter vectors. Back 631717: pSEAP2-Control Vector
631715	pSEAP2-Basic Vector	20 ug	USD $568.00
pSEAP2-Basic is designed to enable the secreted form of human alkaline phosphatase (SEAP) to be used as a reporter gene for analysis of promoter and enhancer sequences. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back pSEAP mammalian reporter vectors pSEAP mammalian reporter vectors. Back 631715: pSEAP2-Basic Vector
631704	Great EscAPe™ SEAP Fluorescence Detection Kit	300 Rxns	USD $527.00
The Great EscAPe SEAP Fluorescence Detection Kit is a complete system for the detection of secreted alkaline phosphatase (SEAP) activity in conditioned media from cultures of mammalian cells expressing this enzyme. The kit contains reagents necessary for the detection of this enzyme activity using the fluorescent substrate 4-methylumbelliferyl phosphate (MUP). Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back SEAP activity is highly specific, with a >20-fold increase in activity SEAP activity is highly specific, with a >20-fold increase in activity. HEK 293 cells were transiently transfected with a promoter construct containing CRE driving the expression of SEAP, or mock-transfected. 12 hr posttransfection, the cell culture media was replaced by media containing either 10 μM forskolin or plain cell culture media and incubated for 7 hr. Forskolin causes an increase in the level of cytosolic cAMP, which in turn activates CRE, driving the expression of SEAP, which is detectable in the media culture supernatant. The samples were assayed using the Great EscAPe SEAP Chemiluminescence Kit 2.0 and its protocol and analyzed on a BD Monolight 3096 Luminometer. Back Comparison of the sensitivities of SEAP and firefly luciferase Comparison of the sensitivities of SEAP and firefly luciferase. Parallel cultures of BHK cells were transiently transfected with the indicated amounts of either pSEAP2-Control Vector or a similar vector expressing luciferase. After 24 hr, SEAP activity was assayed in the appropriate culture media using the Great EscAPe chemiluminescent assay. Similarly, after 24 hr, cell lysates were prepared from the luciferase cultures, and luciferase activity was assayed using a commercial kit. RLU = relative light units. Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription.** Back 631704: Great EscAPe SEAP Fluorescence Detection Kit

Overview

One step assay, for monitoring promoter activity
10-fold higher sensitivity then firefly luciferase
No-cell-lysis protocol
Adaptable from single-tube to high-throughput assays
Dual, live-cell assay includes secreted luciferase as well as SEAP:
monitor two promoters, or use one reporter as a normalization control

More Information

Applications

Time-course studies
Non-invasive reporter gene assays with secreted alkaline phosphatase
Test multiple compounds
Perform downstream experiments with the same cells
Use the dual assay (which includes secreted luciferase, as well a secreted alkaline phosphatase assay) to monitor two promoters
Use the dual assay to monitor one promoter with a built-in normalization control

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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Secreted alkaline phosphatase assays

Fluorescence-based detection

Dual secreted reporter assay—SEAP & luciferase

Notice to purchaser

SEAP activity is highly specific, with a >20-fold increase in activity

Comparison of the sensitivities of SEAP and firefly luciferase

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

631738: Great EscAPe SEAP Chemiluminescence Kit 2.0

Notice to purchaser

SEAP activity is highly specific, with a >20-fold increase in activity

Comparison of the sensitivities of SEAP and firefly luciferase

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

631737: Great EscAPe SEAP Chemiluminescence Kit 2.0

Notice to purchaser

Chemiluminescent detection of protein-protein interactions

SEAP activity is highly specific, with a >20-fold increase in activity

Comparison of the sensitivities of SEAP and firefly luciferase

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

631736: Great EscAPe SEAP Chemiluminescence Kit 2.0

Notice to purchaser

Similar secretion kinetics of Metridia secreted luciferase and SEAP enable accurate comparisons of the relative timing of promoter activity

Monitoring activation of two promoters simultaneously

631734: Ready-To-Glow Dual Secreted Reporter Assay

Notice to purchaser

pSEAP mammalian reporter vectors

631717: pSEAP2-Control Vector

Notice to purchaser

pSEAP mammalian reporter vectors

631715: pSEAP2-Basic Vector

Notice to purchaser

SEAP activity is highly specific, with a >20-fold increase in activity

Comparison of the sensitivities of SEAP and firefly luciferase

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

631704: Great EscAPe SEAP Fluorescence Detection Kit

Overview

More Information

Applications

Additional product information